LOCUS       sequence-482773-        2313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid insert contains the aMFx1 C-terminal tag sequence with the 
            TEV cleavage site for use in Combinatorial Golden Gate Assembly 
            (BsaI restriction sites)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2313)
  AUTHORS   Cleaver A, Luo R, Smith OB, Murphy L, Schwessinger B, Brock J
  TITLE     High-throughput optimisation of protein secretion in yeast via an 
            engineered biosensor.
  JOURNAL   Trends Biotechnol. 2025 Apr;43(4):838-867. doi: 
            10.1016/j.tibtech.2024.11.010. Epub 2024 Dec 13.
  PUBMED    39674781
REFERENCE   2  (bases 1 to 2313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.tibtech.2024.11.010"; journalName: "Trends Biotechnol"; 
            date: "2025-04"; volume: "43"; issue: "4"; pages: "838-867"
COMMENT     Sequence Label: C-term_TEV_aMFx1 (4a)
FEATURES             Location/Qualifiers
     source          1..2313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     464..486
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     483..499
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     483..499
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(766..786)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(908..925)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(908..924)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(917..939)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1026..1128
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1129..1788
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1195..1214)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1809..1903
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      join(2036..2313,1..311)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
       61 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      121 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      181 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      241 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      301 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      361 ttgctcacat gttctttcca gcgacatatg ccatacgccg aactgaactc cacctaatgt
      421 cttggagatt gtcggatagc gcggaatagg attgtgccct ctgagcggat aacaatttca
      481 cacaggaaac agctatgacc atgacacatg catccaccat cgcagactta tcatcactga
      541 gcctccacct agcctaactt caaggctacg tcctattgtt ggccttgtgc atacataaca
      601 tggtttcgta catgtagcgt tcaggagttg tagcaaatcc cacgctgaac gattggggta
      661 acgccttagg attatatcct agcctgtcgg aacacaagca cctttcgtct caggtctcag
      721 ccattaatac attggctgcc ctggctttaa ttgtaaccaa tgccaggatt ggaagtacaa
      781 attttcggat tgagaccgat gagacggcct ctttagtccc gtagaattaa cttttatgcg
      841 accacgctac gtattaggtc aggcggaatg gcacttcctt attccagatg cgtgcggatt
      901 atgcatgact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gcttgtggtg
      961 gatcacagtt attggagact tccttggccg tacgatgggt tttgtttcag gtatcaatga
     1021 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     1081 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1141 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1201 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1261 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1321 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1381 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1441 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1501 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1561 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1621 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1681 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1741 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag
     1801 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     1861 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagtaacaa
     1921 cttatatcgt atggggggac gatttgaagc cccctaccca tttacgctga cttgacggga
     1981 cggcggcttt gttgaataga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2041 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2101 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2161 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2221 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2281 gtggcgataa gtcgtgtctt accgggttgg act
//
